LOCUS       62056_21055        8055 bp DNA     circular SYN 06-OCT-2020
DEFINITION  Cloning vector pTcmKateN-CO 2.0, complete sequence.
ACCESSION   MT180306
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8055)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Characterising ISWI chromatin remodeler in Trypanosoma cruzi
  JOURNAL   Mem. Inst. Oswaldo Cruz 115, e190457 (2020)
  PUBMED    32428081
REFERENCE   2  (bases 1 to 8055)
  AUTHORS   Diaz-Olmos Y, Batista M, Ludwig A, Marchini FK.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz 
            Mader, Curitiba 81350010, Brasil
REFERENCE   3  (bases 1 to 8055)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mem. Inst. 
            Oswaldo Cruz 115, e190457 (2020)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (09-MAR-2020) LACTAS, Fiocruz, Professor Algacyr Munhoz Mader, 
            Curitiba 81350010, Brasil"
FEATURES             Location/Qualifiers
     source          1..8055
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      665..1281
                     /note="Trypanosoma cruzi 18S ribosomal RNA promoter region"
                     /regulatory_class="promoter"
     misc_feature    1294..1798
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511469.50 CDS"
     protein_bind    1816..1940
                     /label=attR1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1965..1995
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     CDS             2049..2705
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     CDS             3050..3352
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     protein_bind    complement(3396..3520)
                     /label=attR2
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             3531..4226
                     /label=mKate2
                     /note="monomeric far-red fluorescent protein (Shcherbo et
                     al., 2009)"
     misc_feature    4244..4664
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.504069.70 CDS"
     CDS             4674..5468
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     misc_feature    5478..5795
                     /note="Trypanosoma cruzi 3' intergenic region of the 
                     TcCLB.511145.40 CDS"
     promoter        complement(5867..5885)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5906..5922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5930..5946)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5954..5984)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5999..6020)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6308..6896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7070..7927)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(7928..8032)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcat
      661 cgatcaccag tttccttgcg gggcttattt ggtgtgttgg atttttaatt ttttgatgat
      721 tcggaggaat tttttgcaat ttttgggtac ccctaaaaaa atttttggag tttccggggt
      781 tcgatccaac ttgtgctttt ttaaccgtcc atttgtgtct agtacatcat ggagcggtat
      841 tctctccggt ggcccaggtg gcgtcggaag gaattggtgt ccctcaggat gtttggtttt
      901 tcgttttttt ttttgcggtc ttttgtttcg catcgccgta tcgtgtgcgg cgcgaccttc
      961 tttttggctc attggtcagt gaggtttata ataaagtgga tcatttgggt ttttggccct
     1021 ttgcggcgca cccgcggaat gggggaggga ttttcatata atgtagtgta cattgtgtgt
     1081 gggtcgcaaa atgtcatgcc gtgtgcaatg tttatttgcg tgctgtgcgt gtccttttgt
     1141 gattccacca cttgtgtgta ttttacctta tcgtctctcc ctgccgttgg ctgtgcgtga
     1201 attgtgcttc gcattcgtgg gaaatcaaaa ggggaatcaa cgtgttcttt tttttcagaa
     1261 ggggagtcaa agcgccgtgt gatcgatcat atggatttgg gtaggatgag aagggagttg
     1321 ctgaggaaaa ggggagggaa tcattaataa ttagaagagt tttttcttct ctctcatttc
     1381 ctgtatctat atatgtatta ggagctgtat tggcaacttg gtggagcatc ggcggggtgt
     1441 ttgcggcgcg gacctctgca aacttccaaa ggtggtaatg ggaatttgaa ataacttgcc
     1501 aatggacatg actcaacaga cgaaagcggt agcaacgata ctatgtaata gctgttgttg
     1561 ttgttatttt tatgtttatt tttattattg atacttcgct tcggtactct ctctgttttt
     1621 ttttttctct cactcattcc ctctttatat atatatatat atatatatat atatatatat
     1681 atcgtacgtg tctttttgtt gttgttctca acttcacgtt ggcttttttt tttttttttt
     1741 tgacttggat tgtatcggac ggcccaataa caataataat taaaataaaa gtggggaaaa
     1801 gcttgcggcc gcatcacaag tttgtacaaa aaagctgaac gagaaacgta aaatgatata
     1861 aatatcaata tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca
     1921 caacatatcc agtcactatg gcggccgcat taggcacccc aggctttaca ctttatgctt
     1981 ccggctcgta taatgtgtgg attttgagtt aggatccgtc gagattttca ggagctaagg
     2041 aagctaaaat ggagaaaaaa atcactggat ataccaccgt tgatatatcc caatggcatc
     2101 gtaaagaaca ttttgaggca tttcagtcag ttgctcaatg tacctataac cagaccgttc
     2161 agctggatat tacggccttt ttaaagaccg taaagaaaaa taagcacaag ttttatccgg
     2221 cctttattca cattcttgcc cgcctgatga atgctcatcc ggaattccgt atggcaatga
     2281 aagacggtga gctggtgata tgggatagtg ttcacccttg ttacaccgtt ttccatgagc
     2341 aaactgaaac gttttcatcg ctctggagtg aataccacga cgatttccgg cagtttctac
     2401 acatatattc gcaagatgtg gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt
     2461 ttattgagaa tatgtttttc gtctcagcca atccctgggt gagtttcacc agttttgatt
     2521 taaacgtggc caatatggac aacttcttcg cccccgtttt caccatgggc aaatattata
     2581 cgcaaggcga caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg
     2641 gcttccatgt cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg
     2701 gggcgtaaac gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     2761 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     2821 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     2881 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     2941 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     3001 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     3061 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
     3121 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     3181 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     3241 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
     3301 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataaatgtc
     3361 aggctccctt atacacagcc agtctgcagg tcgaccatag tgactggata tgttgtgttt
     3421 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
     3481 cattttacgt ttctcgttca gctttcttgt acaaagtggt gatgtctaga atggtgagcg
     3541 agctgattaa ggagaacatg cacatgaagc tgtacatgga gggcaccgtg aacaaccacc
     3601 acttcaagtg cacatccgag ggcgaaggca agccctacga gggcacccag accatgagaa
     3661 tcaaggcggt cgagggcggc cctctcccct tcgccttcga catcctggct accagcttca
     3721 tgtacggcag caaaaccttc atcaaccaca cccagggcat ccccgacttc tttaagcagt
     3781 ccttccccga gggcttcaca tgggagagag tcaccacata cgaagacggg ggcgtgctga
     3841 ccgctaccca ggacaccagc ctccaggacg gctgcctcat ctacaacgtc aagatcagag
     3901 gggtgaactt cccatccaac ggccctgtga tgcagaagaa aacactcggc tgggaggcct
     3961 ccaccgagac cctgtacccc gctgacggcg gcctggaagg cagagccgac atggccctga
     4021 agctcgtggg cgggggccac ctgatctgca acttgaagac cacatacaga tccaagaaac
     4081 ccgctaagaa cctcaagatg cccggcgtct actatgtgga cagaagactg gaaagaatca
     4141 aggaggccga caaagagacc tacgtcgagc agcacgaggt ggctgtggcc agatactgcg
     4201 acctccctag caaactgggg cacagagcat gctaaggcgc gcctatggta gtgaatggat
     4261 atatatatat atatatatat atatatataa ggtgtaaatg aaattagtgg aagagggaat
     4321 tagatgggta gcgcgttgcg tggcgctggg gagtgtctcg ttttatgact tttgtatctc
     4381 tctgtttgag tgtgtgtgtg tgtgtgtgtg tgttttattc tttttttgtt gttgttgttc
     4441 ttgttaaata tttattaatg atttgctttt ccttcttgtg tggcttgaaa ttatttcctc
     4501 tactgtttgt ctccacgtca attttgccca taactcatct tgactttcaa tccttagcga
     4561 gtgttttgtt gtttttggga tagtcgtttt ttgtttgtgt gtgtgtgtgt gtgtgtgtgt
     4621 atttttttct tttttcttct tcttggaggg gctcctacac gtttaccggt actagtatga
     4681 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4741 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4801 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4861 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4921 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4981 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5041 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5101 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5161 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgaatg cccgacggcg
     5221 atgatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5281 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5341 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5401 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5461 agttcttctg actcgagcgg catcggatgc gttttatggt tctgtctttt ttttcccctc
     5521 cttttttttt ttcttttaat ttattttact ttactgcacg gtaccttgta ggaaatgaga
     5581 tgtgtttttt tttttgtttt ttttgttgtt gttgttgttg ttgagatttt tttttttttt
     5641 tttttttttg atcttacgtg tgtcctgtgt tactgagaat atttatgctg cgcatttgga
     5701 gatattcata gttgtttaaa tggacttttt ctgctgctca aacttttgtt ttctgcgtaa
     5761 ttaacaggga aggaaacttt ctaagtcgtt catcagatat cccacttgta tcgtgggcgc
     5821 cacttgaatc gtgggcggcc gcgggcccgg tacccagctt ttgttccctt tagtgagggt
     5881 taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     5941 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     6001 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     6061 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     6121 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     6181 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     6241 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     6301 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     6361 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     6421 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     6481 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     6541 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     6601 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     6661 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     6721 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     6781 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     6841 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     6901 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     6961 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     7021 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     7081 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     7141 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     7201 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     7261 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     7321 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     7381 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     7441 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     7501 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     7561 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     7621 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     7681 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     7741 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     7801 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     7861 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     7921 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     7981 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     8041 cccgaaaagt gccac
//
